An improved apparatus for the optical recording of contraction of single heart cells

An optical system for measuring changes in cell length during unloaded contractions of cardiac myocytes is described. A one-dimensional video image of a cell is obtained every 4 ms with a linear photodiode array, which is aligned with the longitudinal axis of the cell. The circuit used to process the image from the photodiode array has a variety of features to aid in the accurate determination of the distance between the ends of the cell, i.e. the cell length. First, the video image of the cell is divided into two windows, one encompassing the front edge of the cell, the other encompassing the rear edge. Other cells or debris beyond the cell edges are excluded. Changes in the general light level, for example as a result of debris floating above the cell, have little effect because within the windows the background light level is subtracted from the signals before they are processed further. To detect the cell edges, the system determines when the signals within the windows exceed (front edge) or drop below (rear edge) chosen threscholds, which are different for the front and rear edges. The system has memory and it identifies the rear edge of the cell as the last time the signal falls below the threshold; because of this bright spots within the cell are not mistaken for the end of the cell. The system has hysteresis, which enables it to ignore small fluctuations in brightness around the threshold. The system is easy to use, accurate, readily calibrated, and it has good spatial and time resolution (about 0.25 m and 4 ms respectively).     

Biophysical studies of teschen disease virus (Talfan strain)

Summary Studies have been made of the heterogeneity of infectivity and CFA in Teschen virus (Talfan strain) suspensions. Most of the infectivity was contained in two components of densities 1.46 gm./ml. and 1.35 gm./ml. The physical, chemical and immunological properties of these components have been compared. It was possible, however, to convert a large proportion of 1.46 component to 1.35 component by treating the 1.46 component with sodium dodecyl sulphate. This would indicate that the 1.46 component was a complex formed between the infective particles and cellular debris.Further studies on the growth characteristics and electron microscopy of the virus have been made.     

Intratypic antigenic heterogeneity of Coxsackie B viruses

Summary Neutralization tests, employing the cytopathogenic effect in tissue culture tubes, with a variety of homotypic antisera and strains of Coxsackie B viruses often yielded high titers in early readings and low titers in late readings — the break-through phenomenon — and occasionally also low, early-reading titers with heterologous, homotypic sera, which gave high titers with the homologous strains. Of 27 strains of Goxsackie B 1 to B 5, that were tested, 10 showed no break-through tendency while others showed varying degrees of break-through, without reference to any evidence of intratypic antigenic variation. There was a positive correlation between a small number of tissue culture passages away from man or mouse brain and the break-through tendency. Moreover, strains without break-through tendency yielded viral populations with marked break-through properties after a single intracerebral passage in newborn mice, and even after two subsequent tissue culture passages. Plaque-purified progeny exhibited the break-through phenomenon to the same extent as the original, unpurified cultures.The early readings yielded reproducible titers, which could be used for analysis of antigenic variation. Prime antigenic variants, of broader antigenic constitution than their non-prime relatives, were found among the Coxsackie B 2, B 3, and B 4 strains that were tested. These prime strains (e. g., B. V. A. 96- B2; Stevens - B 3, and Burrier or J. V. B. - B 4) were found to be antigenically broader than the prototype strains (Ohio 1 - B 2, Nancy - B 3, Powers or Texas 13 - B 4) generally used for the preparation of diagnostic antisera. The broader antigenicity of the prime variant was also present in plaque-purified progeny.Aided by grants from The National Foundation, Inc.The work reported here was carried out in 1957–1958 during Dr.Wigand's tenure of a fellowship in the Cincinnati Laboratory.     

A cyclin protein modulates mitosis in the budding yeast Saccharomyces cerevisiae

Summary For the budding yeast Saccharomyces cerevisiae the mitotic cell cycle is coordinated with cell mass at the regulatory step start. The threshold amount of cell mass (reflected as a critical size) necessary for start is proportional to nutrient quality. This relationship leads to a transient accumulation of cells at start, termed nutrient modulation, upon enrichment of nutrient conditions. Nutrient enrichment abruptly increases the critical size needed for start, causing the smaller cells, produced in the previous cell cycle, to be delayed at start while growing larger. Here we show that, in S. cerevisiae, a second cell-cycle step, at mitosis, also exhibits nutrient modulation, and is, therefore, another point of cell-cycle regulation. At both mitosis and start, nutrient modulation was found through mutation to be regulated by the activity of the cyclin-related WHI1 (CLN3) gene product.     

Application of the polymerase chain reaction to study the M protein(-like) gene family in beta-hemolytic streptococci

Evaluation of homologous regions of published M protein (emm) gene sequences from group A streptococci (GAS; Streptococcus pyogenes) was used to design three primer pairs for polymerase chain reaction (PCR) and three oligonucleotide probe sequences internal to the amplified products. One set of primers and corresponding probe should detect and lead to amplification of emm(-like) genes of virtually every type (all M), another (SOR^-M) should only amplify emm(-like) genes from GAS negative for serum opacity reaction (SOR) and the third (SOR⁺M) should expand only emm(-like) genes from SOR⁺ GAS. Using the all M primer pair for PCR on the genomic DNA from GAS of 29 different M types as well as from a group C and a group G streptococcal isolate, DNA fragments within the expected size range were amplified in every assay. All PCR products reacted with the all M probe. Related sequences were not detected in genomic DNA of an S. agalactiae and an Enterococcus faecalis isolate. Applying the SOR^-M and SOR⁺M primers to identical assays led to mutually exclusive amplification products. The SOR⁺M and SOR⁺M probes hybridized only to their corresponding products. Exceptions to this exclusivity were the SOR⁺ GAS of M types 3, 8, 27, 34, 42, 67, and 69, which consistently reacted only with the SOR⁺M primer/probe set. Analysis of sequence data from the amplified emm(-like) 2, 3, 18, and 19 genes revealed interesting specific features such as conserved gaps in the C-terminal sequence regions from SOR⁺ and the exceptional SOR^- GAS strains. These data indicate the existence of a subgroup of strains among SOR^- GAS and may advance our understanding of phylogenetic relationship between different serotypes of GAS.     

Effect of the degree of myocardial hypertrophy associated with an experimental heart defect on the resistance to altitude hypoxia

Summary Experimental coarctation of the abdominal aorta with constriction of its lumen to one-third of the original diameter was created in 18 albino rats. Four months later various degrees of myocardial hypertrophy developed in the animals with a relative weight of the heart ranging from 0.0033 to 0.0069. In elevation in the barochamber, the altitude ceiling of the animals with a relative cardiac weight below 0.0040, did not differ from the normal one. The altitude ceiling proved to be considerably decreased in animals with a relative cardiac weight of over 0.0040. Analysis of ECG recorded during the elevation demonstrated that in the animals with a considerable myocardial hypertrophy reduced resistance to the acute high altitude hypoxia depended on the reduction of the functional resistance of the heart.(Presented by Active Member AMN SSSR V. V. Parin) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 55, No. 5, pp. 37–40, May, 1963     

Oncogenic activity of the BK type of human papova virus in inbred rat strains

Summary The oncogenicity of the human polyomavirus BK (BKV) was tested in newborn inbred rats.It was found that the tumor rate was negatively correlated with the levels of T antibody 3 months after inoculation and the frequency of animals with detectable T antibodies 1.5 months after inoculation.By contrast, no influence of viral HI titers on the tumor rates was found. Thymectomy of animals resulted in most experiments in increased tumor rates. Inoculation with BKV of animals later than 24 hours after birth yielded a decrease of tumor rates.The results obtained suggest that T antibody titers present at a critical time after inoculation are associated with low oncogenicity of BKV.The oncogenicity of BKV was comparatively tested in rat strains possessing the allele l or the allele a, respectively. The oncogenicity was significantly higher in rats with the allele l than in rats with the allele a. Rats with the allele l showed lower T antibody response than rats with the allele a.These differences could be explained by the finding that cells of a origin showedin vitro a higher percentage of T antigen bearing cells than did cells of a strain possessing the allele l. In comparison to previous results obtained with BKV inoculated outbred WISTAR rats, the oncogenicity of comparable BKV doses in inbred rats was generally higher and the latency period of tumor manifestation shortened.     

”Reflexes of purpose and freedom” in the comparative physiology of higher nervous activity

The most complex unconditioned reflexes of aim and freedom, discovered by I. P. Pavlov, are compared with the competence drive and the motivation of the resistance to coercion, respectively, described by contemporary ethologists. On the basis of the unconditioned reflex of purpose, conditioned reflexes were developed in which positive emotions arising in connection with the perfection of a skill, irrespective of its pragmatic significance at a given moment, serve as the reinforcement. The unconditioned reflex of freedom is regarded as a phylogenetic precursor of the will, and its acute extinction as the physiological mechanism of hypnosis. It was demonstrated experimentally that the appearance of the state of animal hypnosis (immobilization catatonia) in rabbits is accompanied by the predominance of electrical activity and heat production in the right hemisphere, i. e., by symptoms which are found in hypnosis in man.Translated from Zhurnal Vysshei Nervnoi Deyatel'nosti imeni I. P. Pavlova, Vol. 39, No. 3, pp. 415–420, May–June, 1989.     

Concomitant behavioral and neural inhibition and disinhibition in response to subcortical stimulation

Summary Unit and EEG activity were recorded simultaneously from the same electrode in both acute and chronic cats. In chronic cats, parallels between behavioral and electrophysiological activity were observed. Stimulation of a number of brain sites, including the caudate nucleus and parts of the thalamus, induced an inhibition of unit activity with a concomitant positive (extracellularly recorded) slow wave. The inhibition, which lasted from 150–250 msec, was abbreviated when afferent inputs (thalamocortical or peripheral) were paired with the inhibitory stimuli. When many consecutive pairings were made at a rate of 1/3 to 3 pulses per sec, the abbreviation of inhibition tended to disappear, and, in addition, an inhibitory response to the afferent input alone was often seen. In chronic cats, these electrophysiological observations were paralleled by inhibition and disinhibition of behavioral responses.The authors gratefully acknowledge the technical assistance of: Cora Rucker, David Weber and Marion Smith Baldwin.This work was supported by USPHS Grant MH-07097 and N.A. Buchwald's USPHS Career Development Award 5-K3-GM-3509.     

Endothelial cilia in human aortic atherosclerotic lesions

Summary Primary cilia were present in the endothelial cells of human aortic fatty dots and streaks but not in those of normal intima. They had the features of cilia of the 9+0 axonemal configuration observed in many other cells. A lateral foot process and transitional fibers anchored the ciliary basal body in the cytoplasm, but rootlets were not identified in material examined. Ladder-like configurations interconnected the two centrioles (=diplosome) of control endothelium.The primary cilia of endothelium differed from those of the rudimentary type observed in smooth muscle cells in similar lesions of man, but shared many features with cilia of those present in experimental atherosclerosis in rabbit.Cilia were rarely described in vascular endothelium. It is believed that, to date, they were not reported to occur in normal or pathological arteries in man.It is being stressed that whereas the significance of these unusual organelles remains uncertain, their widespread occurrence may indicate that their role is more important than was believed previously, and they should cease being a curiosity only.Presented-in-part at the Workshop of the American Heart Association: Evolution of the Human Atherosclerotic Plaque, Rockville, Maryland, September 20–23, 1986.Dedicated to Professor Dr. Gotthard Schettler, Department of Internal Medicine, University of Heidelberg, FRG, on the occasion of his 60 birthday (April, 1987).     

Effect of muscular work on the motor function of the stomach and duodenum

Summary The effect of muscle work on the gastric and duodenal motor function was studied in dogs.It was observed that the static stress inhibited the hunger contractions of the stomach and duodenum; the inhibitory effect disappeared as the stress ceased. The static muscle work effect was more pronounced in hunger contractions of the stomach than in those of the duodenum. Under the influence of static stress, the periodic hunger contractions of the stomach varied more extensively than its acid and digestive contractions.Presented by Active Member Acad. Med. Sci. USSR V. N. Chernigovsky     

Zur Frage der Bauart und der mechanischen Beanspruchung mikroskopischer Muskelsehnen

Zusammenfassung Die vorliegende Untersuchung schließt sich an eine Arbeit G. Molliers (1937) an, in der die fächerförmigen Verbreiterungen der Endabschnitte makroskopischer Muskelsehnen erstmalig beschrieben und als Funktionsarchitekturen erkannt worden sind.An Hand eines besonders günstigen Einzelbefundes (Schnittrichtung und Schnittdicke) wird gezeigt, daß G. Molliers Sehnenfächer auch an der periostalen Verankerung der einzelnen mikroskopischen Skeletmuskelfaser verwirklicht ist, so daß er ein allgemein gültiges Konstruktionsprinzip verkörpern dürfte.Die mechanischen Vorteile, die diese Fächerkonstruktion der Sehnenendabschnitte bietet (Ausschaltung gefährlicher Biegebeanspruchung, Vermeidung der Kerbwirkung), werden beschrieben und erläutert.Die Frage, ob die Funktionsarchitektur des Sehnenfächers eine funktionelle Struktur im Sinne W. Roux's, d.h. eine von der Funktion selber verursachte Einrichtung ist, läßt der Verfasser offen.     

Lability in the responses of cells in the auditory cortex of squirrel monkeys to species-specific vocalizations

Summary The activity of 28 cells located mainly in the secondary auditory cortex (A II) of awake squirrel-monkeys, was extracellularly recorded for periods of up to 6 h. Seven different species-specific vocalizations, which were repeatedly presented to the monkey, were used as auditory stimuli. Twenty-six cells responded, at least once, to one or more vocalizations; 22 cells revealed some change in their response (pattern or strength) to at least one vocalization (change in response). Twenty-one cells exhibited a change in the number and/or type of vocalization to which they responded during the recording period (change in selectivity). At some time during the recording period all the responding cells exhibited a change in response and/or a change in selectivity (change in responsiveness). A change in response of a cell to a vocalization did not necessarily exclude a change in selectivity, associated with the same vocalization, later in time and vice-versa. A change in responsiveness to one vocalization was not necessarily correlated with changes in responsiveness to other vocalizations.     

Further observations on the morphogenesis of human rotavirus in MA 104 cells

Summary Human rotavirus KUN strain was cultivated in a fetal rhesus monkey kidney cell line, MA 104 cells. Four types of virus particles in cells infected with KUN strain were clearly identified: nucleoid cores, single-shelled particles, double-shelled particles, and membrane band, enveloped particles. Enveloped particles were found only in the thin sections of infected cells. When first visible, the virus precursors appeared at the ribosome free membrane of rough endoplasmic reticulum (RER), increasing in size while simultaneously being coated with nucleocapsid, inner shell. Single-shelled particles were also synthesized within bundles of filaments of viroplasm in the cytoplasma. During subsequent virus maturation two types of budding processes were observed. Double-shelled particles arising at the RER membrane entered the cisternae of the RER through an exocytosis-like process. In contrast, the enveloped particles developed in the cisternae by being completely enclosed with RER membrane, and later during cytolysis released the single-shelled particles. These enveloped virus particles appeared to be the result of inefficient virus maturation at the last stage of outer capsid formation.With 11 Figures     

Evidence for a direct relationship between mitochondrial genome organization and regeneration ability in hexaploid wheat somatic tissue cultures

Summary Embryogenic and non-embryogenic long-term callus cultures of hexaploid wheat exhibit differences in the organization of their mitochondrial genome. Embryogenic and non-embryogenic fractions of callus cultures initiated from immature embryos of the wheat cultivar Chinese Spring have been isolated and subsequently subcultured. DNA-DNA hybridization experiments using labelled cloned wheat mitochondrial DNA fragments have shown that the mitochondrial DNA organization of embryogenic subcultures derived from embryogenic parts of Chinese Spring calli is closely related to that of the initial Chinese Spring calli, while non-embryogenic subcultures derived from non-embryogenic fragments of Chinese Spring calli exhibit a mitochondrial DNA organization similar to that found in non-embryogenic calli derived from cultivar Aquila. In addition, somatic tissue cultures initiated from three other non-embryogenic wheat cultivars (Talent, Thésée and Capitole) display mitochondrial DNA arrangements similar to those found in cultivar Aquila. These results strongly suggest that, in wheat callus cultures, a particular mitochondrial genome organization is correlated with the ability of cultured cells to regenerate whole plants.Abbreviations mtDNA mitochondrial DNA - ctDNA chloroplast DNA - rRNA ribosomal RNA - kb kilobase pair - cv cultivar - 2,4-D 2,4-dichlorophenoxyacetic acid     

Scope for the “eye — hand” psychophysiological test in screening for cerebrovascular insufficiency

Conclusions 1. During performance of the eye hand PPT the reaction time to input signals at the same time contains information on the trend and level of noise and pulsation of the afferent and efferent pathways of the CNS in man.2. The eye — hand PPT is best performed for detecting subjects with possible CVI, and also to determine the precise degree of its severity.3. Introduction of the eye — hand PPT during mass prophylactic examinations is of economic importance because it enables the early discovery of abnormalities of the workers' health and optimal implementation of the tests in industry characterized by two or three shifts.Riga Medical Institute. Translated from Meditsinskaya Tekhnika, No. 2, pp. 5–7, March–April, 1991.     

Use of B-IV zonal rotor centrifugation as a simple tool for the separation of adeno-associated X7 virus (AAV X7) from helper adenoviruses

Summary The use of zonal rotor centrifugation in the separation of AAV X₇ virus from its helper adenovirus is reported. AAV X₇ virus was perfectly separated from its helper adenovirus. Adeno-associated X₇ virus was found to separate in par tides with an S rate of 72–82 S, and a density in CsCl of 1.30–1.32, and virions with a sedimentation coefficient of 135–147 Svedbergs, with a density of 1.37–1.38.     

Destabilization of herpes simplex virus type 1 virions by local anesthetics,alkaline pH,and calcium depletion

Summary The infectivity of herpes simplex virus type 1 (HSV-1) was found to be markedly reduced by treating virions with the tertiary amine local anesthetics lidocaine, dibucaine and tetracaine. These treatments induced a characteristic shift in the buoyant density of the HSV-1 particles from a light to a heavy population. HSV-1 virions were unstable at alkaline pH, and alkali treatment caused the same shift in buoyant density. Ca²⁺ stabilized the light population. These results suggest that the physicochemical status of the HSV-1 envelope which is sensitive to the treatments described above plays an important role for the integrity of the virion.     

Sodium chloride preference in hypertensive (H) and normotensive (N) rats

Summary Salt consumption was compared in two strains of rats, selected for their disparate proneness (strain H) or resistance (strain N) to Doca-salt hypertension.NaCl intake was similar in H and N rats prior to an following administration of Doca, while their respective blood pressures at the end of this experiment was 178±5 mm Hgvs. 134±3 mm Hg. Thus, disparate responses of the blood pressure to Doca in the two strains cannot be ascribed to differences in salt intake.In another study, salt preference was tested in H and N rats by two-bottle self-selecting technique. Before Doca, saline preference in H rats averaged 60.3±5.8% of total daily fluid consumption,vs 18±4.2% in N rats. Following Doca treatment for 3 weeks the respective values were 96±1.7%vs. 67±6.6%. Thus Doca treatment enhanced salt appetite in both strains, but salt preference remained significantly higher in the H rats.The increased susceptibility to hypertension and the enhanced salt appetite in the H rat, corroborates similar reports in the Okamoto SH rat. In the Brookhaven S rat, however, susceptibility to hypertension is associated with salt avoidance. The conflicting data do not support a unified concept of a genetically determined link between salt appetite and proneness to hypertension.This work was supported in part by a grant from the joint research fund of the Hebrew University-Hadassah Medical School, Jerusalem, and by a grant in aid from Merck Sharpe and Dohme, Research Laboratories.